24 results about "Thrombopoietin" patented technology

The invention provides an anti-human thrombopoietin antibody AF687, a kit and application thereof. The anti-human thrombopoietin antibody disclosed by the invention can specifically recognize epitopes without glycosylation modification of natural TPO protein, and has relatively high affinity for recombinant TPO protein and specific epitopes of the recombinant TPO protein. Meanwhile, the anti-human thrombopoietin antibody disclosed by the invention can accurately detect the content of human thrombopoietin in serum, is not influenced by heterogeneity of natural protein glycosylation sites, can be used for accurately identifying and diagnosing early aplastic anemia and primary immune thrombocytopenia, and has the advantages that compared with an existing detection reagent, the anti-human thrombopoietin antibody is high in sensitivity and high in sensitivity. The false positive rate can be reduced from 13.9% to 2.7%, and a more accurate detection method is provided for differential diagnosis of the primary immune thrombocytopenia.

The invention provides an anti-human thrombopoietin antibody AF692, a kit and application of the anti-human thrombopoietin antibody AF692. The anti-human thrombopoietin antibody disclosed by the invention can specifically recognize epitopes without glycosylation modification of natural TPO protein, and has relatively high affinity for recombinant TPO protein and specific epitopes of the recombinant TPO protein. Meanwhile, the anti-human thrombopoietin antibody disclosed by the invention can be used for accurately detecting the content of human thrombopoietin in serum, is not influenced by natural protein glycosylation site heterogeneity, and can be used for accurately identifying and diagnosing early aplastic anemia and primary immune thrombocytopenia; and a more accurate detection method is provided for differential diagnosis of the primary immune thrombocytopenia.

A hematopoietic stem cell (HSC) proliferation medium includes a basal medium, 0.1 ng / mL to 200.0 ng / mL of thrombopoietin (TPO), 0.1 ng / mL to 200.0 ng / mL of stem cell factor (SCF), 0.1 ng / mL to 20.0 ng / mL of interleukin-3 (IL-3), 0.1 ng / mL to 100.0 ng / mL of interleukin-6 (IL-6), 0.1 ng / mL to 200.0 ng / mL of Flt-3 ligand (FL), 0.1 μg / mL to 20.0 μg / mL of insulin, 0.1 μg / mL to 120.0 μg / mL of transferrin, 0.1 g / L to 3.0 g / L of human serum albumin (HSA), 0.1 μM to 4.0 μM of pyrimido-indole derivative UM729, and 0.1 μM to 2.0 μM of aryl hydrocarbon receptor (AhR) antagonist StemRegenin 1 (SR1). Methods for in vitro proliferation of hematopoietic stem cells (HSCs) and in vitro production of hematopoietic progenitor cells (HPCs) using the HSC proliferation medium are also provided.

The present invention provides a culture medium and a culture method for primary human acute lymphoblastic leukemia cells. The acute lymphoblastic leukemia cell culture medium of the present invention comprises a glutamine additive, non-essential amino acids, recombinant human FLT3 ligand, human interleukin-3, human interleukin-7, human interleukin-2, human stem cell factor, and human thrombopoietin. Using the culture medium and culture method of the present invention, acute lymphoblastic leukemia cells can be cultured with higher amplification efficiency and longer in vitro culture time. The present invention also provides methods and applications for using the culture medium to culture primary human acute lymphoblastic leukemia cells in vitro for drug efficacy evaluation and screening.

The present invention discloses a composition, method and application thereof for improving the quality of in vitro maturation of eggs and the pregnancy rate of embryos after in vitro fertilization. The present invention aims at the problems of low in vitro maturation rate and poor subsequent embryo development potential caused by metabolic disorders during in vitro maturation of oocytes, and provides a composition including two or more regulators in vitamin C, L-carnitine and thrombopoietin to regulate the metabolic mode of oocytes, thereby greatly improving the in vitro maturation rate of oocytes and the subsequent embryo development potential. The present invention not only improves the efficiency of in vitro embryo production of livestock, but also provides a reference for improving the quality of in vitro maturation of human assisted reproductive eggs. The method is simple, safe and efficient, and has great application value and broad application prospects.

The invention discloses a freeze-dried preparation of a pegylated thrombopoietic peptide as well as a preparation method and application of the freeze-dried preparation. The freeze-dried preparation comprises the pegylated thrombopoietic peptide, histidine, glycine, mannitol and cane sugar. The preparation method of the pegylated thrombopoietic peptide comprises the following steps: carrying out coupling treatment on the thrombopoietic peptide and polyethylene glycol to obtain the pegylated thrombopoietic peptide, the thrombopoiesis promoting peptide is prepared from fusion protein; the amino acid sequence of the fusion protein is shown as Xm-(Y-Z) n, X is an insulin glargine mutant, Y is a restriction enzyme cutting site, Z is a thrombopoiesis promoting peptide, m is 1 or 2, and n is 1 or 2. The fusion protein is efficiently expressed in a thallus in the form of an inclusion body, the expressed fusion protein is cut by adopting specific protease, the thrombopoiesis promoting peptide is released, the integrity of the thrombopoiesis promoting peptide can be ensured, and the cutting efficiency is high. The pegylated thrombopoiesis promoting peptide and the preparation thereof can effectively treat and prevent tumor drug related thrombocytopenia, primary immune thrombocytopenia, chronic liver disease related thrombocytopenia and acute radiation syndrome hematopoietic syndrome.

The invention provides a virus vector for hematopoietic stem cells and application of the virus vector. The first aspect of the application provides a lentiviral vector packaging system for hematopoietic stem cells, the lentiviral vector packaging system comprises a recombinant expression vector, and the recombinant expression vector comprises nucleotide sequences encoding thrombopoietin and a fms associated receptor tyrosine kinase 3 ligand. The thrombopoietin and the fms related receptor tyrosine kinase 3 ligand are constructed on the nucleic acid molecule, so that the thrombopoietin and the fms related receptor tyrosine kinase 3 ligand are expressed on the surface of a lentiviral vector and the like; the stemness or immunocompetence of various cells including hematopoietic stem cells can be effectively activated and improved under the condition of lower dosage.

The invention relates to the technical field of stem cell culture. The invention provides a stem cell three-dimensional culture medium and application thereof. The culture medium comprises a basic culture medium and an additive, the additives comprise collagen, Pluronic F-68, hyaluronic acid, folic acid, basic fibroblast growth factors, linoleic acid, arachidonic acid, platelet-derived growth factors, epidermal growth factors, insulin-like growth factor-1, SCF stem cell factors, thrombopoietin, vitamin C, vitamin E, vitamin B1, vitamin B2 and vitamin B6. The culture medium provided by the invention does not need a three-dimensional carrier, and stem cells can be directly cultured in a suspension culture mode. Meanwhile, cell adherence can be effectively prevented, collagenase is not needed in subsequent treatment, the in-vitro culture amplification speed and the cell survival rate are further increased, and the method is suitable for large-scale production and preparation of stem cells.

Embodiments of the disclosure include systems, methods, and compositions for producing megakaryocytes and platelets for recipient individuals in need thereof. In certain embodiments the megakaryocytes and platelets may be produced following co-culture of MSCs and CD34+ cells in media comprising stem cell factor, thrombopoietin, IL-6, IL-3, FLT3-ligand, and an inhibitor of Rho associated coiled – coil containing protein kinase (ROCK).

The invention provides application of thrombopoietin in preparation of CAR-T (Chimeric Antigen Receptor-T) cells and a preparation method of the CAR-T cells. The preparation method of the CAR-T can effectively shorten the production cycle, is simple and convenient to operate, and has a wide industrial application prospect.

The invention provides a cell population and a method for obtaining the same. A cell population according to the present invention is obtained by culturing mononuclear cells derived from bone marrow, umbilical cord blood or peripheral blood in a culture medium containing serum and four or less factors selected from the group consisting of stem cell factors, interleukin 6, FMS-like tyrosine kinase 3 ligand, thrombopoietin, and vascular endothelial cell growth factors.

The invention discloses a complete set composition for obtaining NK cells from umbilical cord blood and a method for culturing the NK cells, and belongs to the technical field of biology. The invention provides a complete set composition for obtaining NK cells from umbilical cord blood, and the complete set composition comprises a composition 2, and the composition 2 is composed of IL-2, IL-15, IL-18, IL-27 and IL-21. The complete set of composition further comprises a composition 1, and the composition 1 is composed of thrombopoietin, stem cell factors and IL-2. The culture medium has the advantages that compared with the prior art, the culture medium prepared from the whole set of composition can remarkably improve the purity of NK cells obtained from umbilical cord blood, and the NK phenotype proportion reaches 85% or above.

The present invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for detecting and analyzing the peptide map of thrombopoietin-mimetic peptides, comprising the following steps: (1) The thrombopoietin-mimetic peptides are subjected to buffer system replacement by ultrafiltration centrifugation; (2) Trypsin is used to enzymatically digest the thrombopoietin-mimetic peptides; (3) Reverse-phase high-performance liquid chromatography is used to detect and analyze the enzymatically digested peptide segments. The beneficial effects of the present invention are as follows: In the present invention, sample concentration and buffer replacement are carried out through an ultrafiltration tube, and the sample after enzymatic digestion has high purity and can be directly detected by reverse-phase high-performance liquid chromatography. Therefore, the method for detecting and analyzing the peptide map of thrombopoietin-mimetic peptides is simple to operate, improving the detection efficiency of thrombopoietin-mimetic peptide drugs; in the present invention, by controlling the chromatographic conditions, the accuracy and precision of high-performance liquid chromatography detection are improved, enabling the quality analysis and control of thrombopoietin-mimetic peptide drugs.

A method for preparing an immune cell differentiated from a stem cell is provided. The method includes: (a) forming a coating containing a matrix on a surface; and (b) culturing a stem cell on the coating in the presence of a first cytokine combination to differentiate the stem cell into an immune cell, wherein the immune cell includes a γδ T cell (gamma delta T cell, GDT cell). The matrix includes: vascular cell adhesion molecule (VCAM); intercellular adhesion molecule (ICAM); and delta-like ligand 4 (DLL4). The first cytokine combination includes: stem cell factor (SCF); thrombopoietin (TPO); Fms-like tyrosine kinase 3 ligand (Flt3L); and interleukin-7 (IL-7). Moreover, a source of the stem cell includes umbilical cord blood or an induced pluripotent stem cell (iPSC), and the stem cell is CD34 positive.

The invention discloses a pharmaceutical composition for treating and curing acute radiation sickness, which comprises idebegrasbine alpha and a thrombopoiesis promoting drug (ropursbine or rhTPO), according to the application disclosed by the invention, for the first time, the fact that the combination of the idebesisterone alpha and the thrombopoiesis promoting drug (ropsisterone or rhTPO) has a remarkable synergistic effect on the treatment of the acute radiation sickness is found, and an effective drug combination strategy is provided for the treatment of the acute radiation sickness; the compound has a wide application prospect in the technical field of development of acute radiation sickness prevention and treatment medicines.

The present invention relates to isolated and modified thrombopoietin polypeptides, uses thereof in expanding stem cells and producing platelets, and therapeutic applications of such modified thrombopoietin polypeptides, stem cells and platelets.

The invention discloses an adeno-associated virus AAV vector. The adeno-associated virus AAV vector comprises a cytokine gene, the cell factor is selected from any one or more than two of the following components: human interleukin 2 (IL2), human interleukin 3 (IL3), human interleukin 6 (IL6), human interleukin 15 (IL15), human thrombopoietin (THPO), human stem cell factor (SCF) and human granulocyte-macrophage colony stimulating factor (GM-CSF). The invention also discloses an AAV vector combination comprising any two or more than three AAV vectors mentioned above. The invention also discloses an application of the AAV vector and a combination of the AAV vector in construction of immune system humanized non-human animals. The invention further discloses a method for constructing the immune system humanized non-human animal.

Provided is a method of inducing the proliferation and / or differentiation of induced pluripotent cells, embryonic stem cells, hematopoietic stem / progenitor cells and megakaryocytes into platelets, comprising culturing cells with a culture medium containing TPO (thrombopoietin), a SCF (stem cell factor), Flt3 (Fms-related tyrosine kinase 3 ligand), IL-3 (interleukin-3), IL-6 (interleukin-6), an AHR antagonist and PVA (polyvinyl alcohol). Also provided are a culture medium for inducing cell proliferation and / or differentiation into platelets, a composition, a culture platform and the use thereof.

The invention belongs to the technical field of biological genetic engineering, and particularly relates to a canine TPO recombinant protein, a mutant, a fusion protein and application thereof. A recombinant protein is obtained by screening a high-expression signal peptide and replacing a natural signal peptide in canine TPO, protein expression can be efficiently induced, mutant protein with higher expression quantity and biological activity is obtained by performing amino acid site mutation on a mature peptide cTPO in canine TPO, and the canine TPO recombinant protein and a mutant thereof are fused with truncated CTP, so that the canine TPO recombinant protein is obtained. The expression quantity and activity of the fusion protein are remarkably improved, an amino acid sequence of the fusion protein is subjected to codon optimization modification and then is expressed by using an efficient CHO expression system, and the problems that in the prior art, dog-derived thrombopoietin is short in half-life period and insufficient in stability, and heterologous protein has immunogenicity worry are solved. The canine TPO fusion protein can be effectively applied to preparation of drugs for treating canine primary or secondary thrombocytopenia.

Provided are a nucleic acid encoding thrombopoietin or a precursor thereof, and a composition and kit comprising same. Further provided is a use of the nucleic acid, the composition and the kit in treatment of thrombocytopenia.

This application provides a viral vector for hematopoietic stem cells and its application. A first aspect of this application proposes a lentiviral vector packaging system for hematopoietic stem cells, the lentiviral vector packaging system comprising a recombinant expression vector, the recombinant expression vector comprising nucleotide sequences encoding thrombopoietin and FMS-associated receptor tyrosine kinase 3 ligand. This invention, by constructing thrombopoietin and FMS-associated receptor tyrosine kinase 3 ligand onto nucleic acid molecules, thereby enabling their expression on the surface of a lentiviral vector, shows that compared to direct addition, it can effectively activate and enhance the stemness or immune activity of various cells, including hematopoietic stem cells, at lower dosages.

Provided herein are methods, compounds, and compositions for safely reducing thrombopoietin in a cell or an individual. Such methods, compounds, and compositions maintain platelet count within a safe hemostatic range. Such methods, compounds, and compositions are useful to safely treat, prevent, or ameliorate a disease that can benefit from platelet count reduction in an individual. Such methods, compounds, and compositions are useful for treating or preventing diseases in which thrombopoietin contributes to the presence or activation of platelets contributes to and promotes disease initiation or progression, and / or adversely affects disease outcome.

The present disclosure discloses a composition, method and application thereof for improving the quality of in vitro maturation of oocytes and the pregnancy rate after in vitro produced embryos transfer. The present disclosure aims at the problems of low maturation rate and poor subsequent embryo development potential caused by metabolic disorders during in vitro maturation of oocytes, and provides a composition including two or more regulators in vitamin C, L-carnitine and thrombopoietin to regulate the metabolism of oocytes, thereby greatly improving the in vitro maturation rate of oocytes and the subsequent embryo development potential. The present disclosure not only improves the efficiency of in vitro embryo production of livestock, but also provides a reference for improving the efficiency of human assisted reproductive technology. The method is simple, safe and efficient, and has great application value and broad application prospects.

Provided herein are methods, compounds, and compositions for safely reducing thrombopoietin in a cell or an individual. Such methods, compounds, and compositions maintain platelet count within a safe hemostatic range. Such methods, compounds, and compositions are useful to safely treat, prevent, or ameliorate a disease that can benefit from platelet count reduction in an individual. Such methods, compounds, and compositions are useful for treating or preventing diseases in which thrombopoietin contributes to the presence or activation of platelets contributes to and promotes disease initiation or progression, and / or adversely affects disease outcome.